毕赤酵母赖氨酰氧化酶中醌辅因子的鉴定。

Identification of the quinone cofactor in a lysyl oxidase from Pichia pastoris.

作者信息

Dove J E, Smith A J, Kuchar J, Brown D E, Dooley D M, Klinman J P

机构信息

Department of Chemistry, University of California, Berkeley 94720, USA.

出版信息

FEBS Lett. 1996 Dec 2;398(2-3):231-4. doi: 10.1016/s0014-5793(96)01245-8.

DOI:10.1016/s0014-5793(96)01245-8

PMID:8977113

Abstract

A copper amine oxidase from Pichia pastoris is the only known non-mammalian lysyl oxidase [Tur, S.S. and Lerch, K. (1988) FEBS Lett. 238, 74-76]. Recently, the cofactor in mammalian lysyl oxidase has been identified as a novel lysine tyrosylquinone moiety [Wang, S.X., Mure, M., Medzihradszky, K.F., Burlingame, A.L., Brown, D.E., Dooley, D.M., Smith, A.J., Kagan, H.M. and Klinman, J.P. (1996) Science 273, 1078-1084]. In order to identify the cofactor in P. pastoris lysyl oxidase, we have isolated the phenylhydrazone-derivative of the active-site peptide. This peptide has the active-site sequence conserved among topa quinone containing amine oxidases. The resonance Raman spectra of the phenylhydrazone derivatives of the enzyme, active-site peptide, and a topa quinone model compound are essentially identical. Collectively, these results establish that P. pastoris lysyl oxidase is a topa quinone enzyme.

摘要

来自毕赤酵母的一种铜胺氧化酶是唯一已知的非哺乳动物赖氨酰氧化酶[Tur, S.S.和Lerch, K. (1988) 《欧洲生物化学学会联合会快报》238, 74 - 76]。最近，已确定哺乳动物赖氨酰氧化酶中的辅因子是一种新型的赖氨酸酪氨酰醌部分[Wang, S.X., Mure, M., Medzihradszky, K.F., Burlingame, A.L., Brown, D.E., Dooley, D.M., Smith, A.J., Kagan, H.M.和Klinman, J.P. (1996) 《科学》273, 1078 - 1084]。为了确定毕赤酵母赖氨酰氧化酶中的辅因子，我们分离出了活性位点肽的苯腙衍生物。该肽具有在含topa醌的胺氧化酶中保守的活性位点序列。该酶、活性位点肽和一种topa醌模型化合物的苯腙衍生物的共振拉曼光谱基本相同。总体而言，这些结果表明毕赤酵母赖氨酰氧化酶是一种topa醌酶。